驱虫药尼氯硝唑对结肠癌中 S100A4 介导的转移进展的新作用。

Novel effect of antihelminthic Niclosamide on S100A4-mediated metastatic progression in colon cancer.

机构信息

Max-Delbrück-Center for Molecular Medicine, Berlin, Germany.

出版信息

J Natl Cancer Inst. 2011 Jul 6;103(13):1018-36. doi: 10.1093/jnci/djr190. Epub 2011 Jun 17.

DOI:10.1093/jnci/djr190

PMID:21685359

Abstract

BACKGROUND

Metastasis formation in colon cancer severely reduces the survival rate in patients. S100A4, a calcium-binding protein, is implicated in promoting metastasis formation in colon cancer.

METHODS

To identify a transcription inhibitor of S100A4, high-throughput screening of 1280 pharmacologically active compounds was performed using a human colon cancer cell line expressing a S100A4 promoter-driven luciferase (LUC) reporter gene construct (HCT116-S1004p-LUC). Niclosamide, an antihelminthic agent, was identified as a potential candidate. Colon cancer cell lines (HCT116, SW620, LS174T, SW480, and DLD-1) were treated with 1 μM niclosamide to analyze the effect on S100A4 mRNA and protein expression by quantitative reverse transcription-polymerase chain reaction and immunoblot assays, and effects on cell migration, invasion, proliferation, and colony formation were also assessed in vitro. The effect of niclosamide on liver metastasis was assessed in a xenograft mouse model of human colon cancer (n = 8 mice) by in vivo imaging. The long-term effect of niclosamide on metastasis formation after discontinued treatment was quantified by scoring, and overall survival (n = 12 mice) was analyzed by Kaplan-Meier method after discontinuation of treatment. All statistical tests were two-sided.

RESULTS

Reduced S100A4 mRNA and protein expression, and inhibited cell migration, invasion, proliferation, and colony formation were observed in niclosamide-treated colon cancer cells in vitro. In vivo imaging of niclosamide-treated mice showed reduced liver metastasis compared with solvent-treated control mice (n = 4 mice per group). Compared with the control group, discontinuation of treatment for 26 days showed reduced liver metastasis formation in mice (n = 6 mice per group) (control vs discontinuous treatment, mean metastasis score = 100% vs 34.9%, mean difference = 65.1%; 95% confidence interval [CI] = 18.4% to 111.9%, P < .01) and increased overall survival (n = 6 mice per group; control vs discontinuous treatment, median survival = 24 vs 46.5 days, ratio = 0.52, 95% CI = 0.19 to 0.84, P = .001).

CONCLUSION

Niclosamide inhibits S100A4-induced metastasis formation in a mouse model of colon cancer and has therapeutic potential.

摘要

背景

结肠癌转移的形成严重降低了患者的生存率。S100A4 是一种钙结合蛋白，它被认为可以促进结肠癌的转移形成。

方法

为了鉴定 S100A4 的转录抑制剂，使用表达 S100A4 启动子驱动的荧光素酶（LUC）报告基因构建体（HCT116-S1004p-LUC）的人结肠癌细胞系进行了 1280 种药理学活性化合物的高通量筛选。发现驱虫剂尼氯硝唑是一种潜在的候选药物。用 1 μM 尼氯硝唑处理结肠癌细胞系（HCT116、SW620、LS174T、SW480 和 DLD-1），通过定量逆转录聚合酶链反应和免疫印迹分析来分析对 S100A4 mRNA 和蛋白表达的影响，还评估了体外细胞迁移、侵袭、增殖和集落形成的影响。通过体内成像评估了尼氯硝唑在人结肠癌异种移植小鼠模型中的肝转移抑制作用（n = 8 只小鼠）。通过评分量化了尼氯硝唑停药后对转移形成的长期影响，并通过 Kaplan-Meier 方法分析了停药后（n = 12 只小鼠）的总生存情况。所有统计检验均为双侧检验。

结果

在体外，尼氯硝唑处理的结肠癌细胞中 S100A4 mRNA 和蛋白表达减少，细胞迁移、侵袭、增殖和集落形成受到抑制。与溶剂处理的对照组小鼠相比，尼氯硝唑处理的小鼠体内成像显示肝转移减少（每组 n = 4 只小鼠）。与对照组相比，停药 26 天后，小鼠肝转移形成减少（每组 n = 6 只小鼠）（对照组与间断治疗组，平均转移评分= 100% vs 34.9%，平均差异= 65.1%；95%置信区间[CI] = 18.4%至 111.9%，P <.01），总生存期延长（每组 n = 6 只小鼠；对照组与间断治疗组，中位生存= 24 天 vs 46.5 天，比值= 0.52，95%CI = 0.19 至 0.84，P =.001）。